1. Field of the Invention
The present invention relates to a method for massive culture of Dinophysis acuminata, the marine dinoflagellate causing diarrhetic shellfish poisoning, and methods for extracting, isolating and purifying the shellfish toxin pectenotoxin-2 from the cultured Dinophysis acuminata. 
2. Description of Related Art
Dinophysis sp. belonging to dinoflagellates causing red tide is known to produce pectenotoxin (PTX) which is one of diarrhetic shellfish toxins. Once this organism causes red tide in the ocean, it brings huge damage on marine shellfish aquaculture. The major toxins produced by this living thing are okadaic acid, pectenotoxin, dinophysistoxin and yessotoxin, etc. It has been recently reported that pectenotoxin induces apoptosis selectively of cancer cells. So, expectation of medical or pharmaceutical usability of this material is increasing. In addition, pectenotoxin can be used as a standard material for toxin especially when red tide occurs by this living thing in the ocean. Although pectenotoxin has a wide spectrum of application, studies on the material have been limited for a long time because massive-culture of Dinophysis was unsuccessful and thus pectenotoxin could only picked up from a limited area.
Dinophysis acuminate, the marine dinoflagellate is a causing organism of diarrhetic shellfish poisoning which could not be cultured successfully in a lab until the year of 2006. The culture of Dinophysis acuminate was first succeeded by supplying laboratory culture of Myrionecta rubra, a marine ciliate, as feed in 2006 (Park et al. 2006. First successful culture of the marine dinoflagellate Dinophysis acuminate. Aquat. Microb. Ecol. 45, 101-106). Since then, success of culture of Dinophysis acuminate or other Dinophysis species using Myrionecta rubra has been reported from time to time.
However, the marine dinoflagellate, Dinophysis acuminate has been cultured only in a laboratory scale. So, the culture amount of Dinophysis acuminate is always not enough for the study of pectenotoxin, considering the increasing medical or pharmaceutical usability of the material. Therefore, a method for massive culture of Dinophysis acuminate is urgently requested for the study of pectenotoxin.
Based on the early method for culture of a small amount of Dinophysis acuminate, which was succeeded in a lab, the present inventors designed a simple water bath for massive culture of 500 liter and tried to establish a method for culture of the organism and a method for extracting and separating pectenotoxin, the diarrhetic shellfish toxin contained in the organism.